De novo design of a redox-active minimal rubredoxin mimic
De novo design of a redox-active minimal rubredoxin mimic
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DOI：
10.1021/ja050553f
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发表时间：
2005-04-27
影响因子：
15
通讯作者：
DeGrado, WF
中科院分区：
文献类型：
--
作者：
Nanda, V;Rosenblatt, MM;DeGrado, WF
金属蛋白中的金属结合位点经常出现在二级结构元素的界面处，这使得天然蛋白质的逆结构分析和结合金属离子辅助因子的螺旋束的从头设计成为可能。然而，尽管天然金属蛋白中经常出现β构象，但含有β结构的金属蛋白的设计还不太成熟。在这里，我们描述了 β-蛋白 RM1 的设计和构建，该蛋白形成稳定的氧化还原活性 4-Cys 硫醇盐 Fe(II/III) 位点，类似于红氧还蛋白的活性位点。该蛋白质在存在或不存在金属离子的情况下折叠成 β 结构，并结合 Fe(II/III) 形成氧化还原活性位点，即使在有氧环境中，该位点也能稳定地重复氧化和还原循环。
Metal-binding sites in metalloproteins frequently occur at the interfaces of elements of secondary structure, which has enabled the retrostructural analysis of natural proteins and the de novo design of helical bundles that bind metal ion cofactors. However, the design of metalloproteins containing β-structure is less well developed, despite the frequent occurrence of β-conformations in natural metalloproteins. Here, we describe the design and construction of a β-protein, RM1, that forms a stable, redox-active 4-Cys thiolate Fe(II/III) site analogous to the active site of rubredoxin. The protein folds into a β-structure in the presence and absence of metal ions and binds Fe(II/III) to form a redox-active site that is stable to repeated cycles of oxidation and reduction, even in an aerobic environment.